PMID- 12946945 OWN - NLM STAT- MEDLINE DCOM- 20030930 LR - 20131121 IS - 1524-4571 (Electronic) IS - 0009-7330 (Linking) VI - 93 IP - 6 DP - 2003 Sep 19 TI - Fas and Fas-associated death domain protein regulate monocyte chemoattractant protein-1 expression by human smooth muscle cells through caspase- and calpain-dependent release of interleukin-1alpha. PG - 515-22 AB - We previously reported that treatment of human vascular smooth muscle cells (SMCs) with proapoptotic stimuli, including Fas ligand plus cycloheximide (FasL/Chx), or overexpression of Fas-associated death domain protein (FADD) result in increased expression of monocyte chemoattractant protein-1 (MCP-1) and other proinflammatory genes. In this study, we demonstrate that Fas/FADD-induced MCP-1 upregulation is driven by an autocrine/paracrine signaling loop in which interleukin (IL)-1alpha synthesis and release are activated through caspase- and calpain-dependent processes. Untreated SMCs contain very little IL-1alpha protein or transcript. Both were increased greatly in response to Fas/FADD activation, primarily through an autocrine/paracrine pathway in which secreted IL-1alpha stimulated additional IL-1alpha synthesis and release. Caspase 8 (Csp8) activity increased in response to FasL/Chx treatment, and Csp8 inhibitors markedly reduced IL-1alpha release and MCP-1 upregulation. In contrast, Csp8 activity was not significantly increased in response to FADD overexpression and caspase inhibitors did not effect FADD-induced MCP-1 upregulation. Both FasL/Chx treatment and FADD overexpression increased the activity of calpains. Calpain inhibitors reduced IL-1alpha release and MCP-1 upregulation in both FADD-overexpressing SMCs and FasL/Chx-treated SMCs without blocking Csp8 activity. This indicates that calpains are not required for activation of caspases and that caspase activation is not sufficient for IL-1alpha release and MCP-1 upregulation. These data suggest that calpains play a dominant role in Fas/FADD-induced IL-1alpha release and MCP-1 upregulation and that caspase activation may function to amplify the effects of calpain activation. FAU - Schaub, Friedemann J AU - Schaub FJ AD - Department of Pathology, University of Washington, Box 357470, Seattle, Wash 98195-7470, USA. FAU - Liles, W Conrad AU - Liles WC FAU - Ferri, Nicola AU - Ferri N FAU - Sayson, Kirsten AU - Sayson K FAU - Seifert, Ronald A AU - Seifert RA FAU - Bowen-Pope, Daniel F AU - Bowen-Pope DF LA - eng GR - HL62995/HL/NHLBI NIH HHS/United States GR - HL69066/HL/NHLBI NIH HHS/United States PT - Journal Article PT - Research Support, U.S. Gov't, P.H.S. DEP - 20030828 PL - United States TA - Circ Res JT - Circulation research JID - 0047103 RN - 0 (Adaptor Proteins, Signal Transducing) RN - 0 (Carrier Proteins) RN - 0 (Chemokine CCL2) RN - 0 (FADD protein, human) RN - 0 (FASLG protein, human) RN - 0 (Fas Ligand Protein) RN - 0 (Fas-Associated Death Domain Protein) RN - 0 (Interleukin-1) RN - 0 (Membrane Glycoproteins) RN - 98600C0908 (Cycloheximide) RN - EC 3.4.22.- (Calpain) RN - EC 3.4.22.- (Caspases) SB - IM MH - *Adaptor Proteins, Signal Transducing MH - Calpain/metabolism/*physiology MH - Carrier Proteins/*metabolism MH - Caspases/metabolism/*physiology MH - Cells, Cultured MH - Chemokine CCL2/*biosynthesis/genetics MH - Cycloheximide/pharmacology MH - Fas Ligand Protein MH - Fas-Associated Death Domain Protein MH - Gene Expression Regulation MH - Humans MH - Interleukin-1/*biosynthesis/genetics/physiology MH - Membrane Glycoproteins/*pharmacology MH - Muscle, Smooth, Vascular/drug effects/enzymology/*immunology MH - Signal Transduction MH - Transcription, Genetic/drug effects MH - Up-Regulation EDAT- 2003/08/30 05:00 MHDA- 2003/10/01 05:00 CRDT- 2003/08/30 05:00 PHST- 2003/08/30 05:00 [pubmed] PHST- 2003/10/01 05:00 [medline] PHST- 2003/08/30 05:00 [entrez] AID - 01.RES.0000093205.42313.7C [pii] AID - 10.1161/01.RES.0000093205.42313.7C [doi] PST - ppublish SO - Circ Res. 2003 Sep 19;93(6):515-22. doi: 10.1161/01.RES.0000093205.42313.7C. Epub 2003 Aug 28.